Summary
The neuropeptide tyrosine precursor (pre-pro-NPY) messenger RNA (mRNA) has been localised in formaldchyde-fixed human phaeochromocytoma tissue using a sensitive in situ hybridisation procedure and a novel single-stranded cDNA probe. The reaction product was revealed by avidin-biotin-peroxidase complex and streptavidin-gold complex with silver enhancement. This technique may be applied for the determination of biosynthetic activity of endocrine and neuronal cell bodies. This is largely due to its rapidity by comparison with conventional autoradiographic procedures, to the permanence of the reaction product and to the sensitivity of the visualisation steps.
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Adrian TE, Allen JM, Terenghi G, Bacarese-Hamilton AJ, Brown MJ, Polak JM, Bloom SR (1983) Neuropeptide Y in phaeochromocytomas and ganglioneuroblastomas. Lancet 2:540–543
Allen YS, Adrian TE, Allen JM, Tatemoto K, Crow TJ, Bloom SR, Polak JM (1983) Neuropeptide Y distribution in rat brain. Science 221:877–879
Bonnard C, Papermaster DS and Kraehenbuhl J-P (1984) The streptavidin-biotin bridge technique: application in light and electron microscope immunocytochemistry. In: Polak JM, Varndell JM (eds) Immunolabelling for electron microscopy. Elsevier, Amsterdam, pp 95–111
Brigati DJ, Myerson D, Leary JJ, Spalholz B, Travis SZ, Fong CKY, Hsiung GD, Ward DC (1983) Detection of viral genomes in cultured cells and paraffin embedded tissue sections using biotin-labelled hybridisation probes. Virology 126:32–50
Coghlan J, Penschow JD, Hudson PJ, Niall HD (1984) Hybridization histochemistry: use of recombinant DNA for tissue localization of specific mRNA populations. Clin Exp Hypertension Theory Practice A6:63–78
Danscher G, Nörgaard JOR (1983) Light microscopic visualisation of colloidal gold on resin-embedded tissue. J Histochem Cytochem 31:1394–1398
Gee CE, Chen C-LC, Roberts JL, Thompson R, Watson SJ (1983) Identification of proopiomelanocortin neurones in rat hypothalamus by cDNA-mRNA in situ hybridization. Nature 306:374–376
Gee CE, Roberts JL (1983) In situ hybridization histochemistry: a technique for the study of gene expression in single cells. DNA 2:157–163
Gu J, Polak JM, Adrian TE, Allen JM, Tatemoto K, Bloom SR (1983) Neuropeptide tyrosine (NPY): a major new cardiac neuropeptide. Lancet 1:1008–1010
Heidecker G, Messing J, Groneborn B (1980) A versatile primer for DNA sequencing in the M13mp2 cloning system. Gene 10:69–73
Holgate C, Jackson P, Cowen PN, Bird CC (1983) Immunogoldsilver staining: new method of immunostaining with enhanced sensitivity. J Histochem Cytochem 31:938–944
Hsu S-M, Raine L, Fanger H (1981) Use of avidin-biotin-peroxidase complex (ABC) in immunoperoxidase techniques: a comparison between ABC and unlabeled antibody (PAP) procedures. J Histochem Cytochem 29:577–580
Huang W-M, Gibson SJ, Facer P, Gu J, Polak JM (1983) Improved section adhesion for immunocytochemistry using high molecular weight polymers of L-lysine as a slide coating. Histochemistry 77:275–279
Hudson P, Penschow J, Shine J, Ryan G, Niall H, Coghlan J (1981) Hybridization histochemistry: use of recombinant DNA as a “homing probe” for tissue localization of specific mRNA populations. Endocrinology 108:353–356
Lundberg JM, Terenius L, Hökfelt T, Martling CR, Tatemoto K, Mutt V, Polak JM, Bloom SR, Goldstein M (1982) Neuropeptide Y (NPY)-like immunoreactivity in peripheral noradrenergic neurons and effects of NPY on sympathetic function. Acta Physiol Scand 116:477–480
Maxam A, Gilbert W (1980) Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol 65:499–560
Minth CD, Bloom SR, Polak JM, Dixon JE (1984) Cloning and DNA sequence of a human cDNA encoding neuropeptide tyrosine (NPY). Proc Natl Acad Sci USA 81:4577–4581
Minth CD, Taylor WL, Magazin M, Tavianini MA, Collier K, Weith HL, Dixon JE (1982) The structure of cloned DNA complementary to catfish pancreatic somatostatin-14 messenger RNA. J Biol Chem 257:10372–10377
Polak JM, Bloom SR (1984) Neuroendocrine tumours. Brit J Hosp Med (in press)
Sabate MI, Carlei F, Bloom SR, Polak JM (1984) Endocrine tumours of the gut and pancreas. In: Polak JM, Bloom SR (eds) Endocrine tumours — the pathobiology of regulatory peptideproducing tumours. Churchill-Livingstone, Edinburgh (in press)
Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467
Schmechel DE, Marangos PJ, Brightman MW (1979) Neuron specific enolase is a marker for peripheral and central neuroendocrine cells. Nature 276:834–836
Singer RH, Ward DC (1982) Actin gene expression visualized in chicken muscle tissue culture by using in situ hybridization with a biotinated nucleotide analog. Proc Natl Acad Sci USA 79:7331–7335
Springall DR, Hacker GW, Grimelius L, Polak JM (1984) The potential of the immunogold-silver staining method for paraffin sections. Histochemistry 81:603–608
Tapia FJ, Polak JM, Barbosa AJA, Bloom SR, Marangos PJ, Dermody CJ, Pearse AGE (1981) Neuron-specific enolase is produced by neuroendocrine tumours. Lancet 1:808–811
Tatemoto K (1982) Neuropeptide Y: complete amino acid sequence of the brain peptide. Proc Natl Acad Sci USA 79:5485–5489
Varndell IM, Polak JM, Allen JM, Terenghi G, Bloom SR (1984) Neuropeptide tyrosine (NPY) immunoreactivity in norepinephrine-containing cells and nerves of the mammalian adrenal gland. Endocrinology 114:1460–1462
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Varndell, I.M., Polak, J.M., Sikri, K.L. et al. Visualisation of messenger RNA directing peptide synthesis by in situ hybridisation using a novel single-stranded cDNA probe potential for the investigation of gene expression and endocrine cell activity. Histochemistry 81, 597–601 (1984). https://doi.org/10.1007/BF00489541
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DOI: https://doi.org/10.1007/BF00489541